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简介：Genestellcellswhattodo,forexample,whentorepairDNAmistakesorwhentodie,andcanbeturnedonorofflikealightswitch.Knowingwhichgenesareswitchedon,orexpressed,isimportantforthetreatmentandmonitoringofdisease.Now,forthefirsttime,Researchersinthelaboratoryof

简介：Objective:TostudytheimmunologicalmechanismsofCondylomaacuminata(CA)throughinvestigatingTlymphocytesubsetlevelsandcytokineprofileintheperipheralbloodofpatientswithCondylomaAcuminata.Methods:Tricolorandbicolorimmunofluorescentstainingantibodyofcellsurfaceantigenandintracel-lularIL-2,IL-4,IL-12,IFN-γinCD4^+andCD8^+T-lymphocytesfrom20patientswithCAwereperformedandfollowedbyflowcytometry.Results:ThenumberofCD3^+T,CD4^+T-lymphocytescellsandCD4^+/CD8^+Tcellsratioweresignificantlydecreased(P<0.01)inpatientswithCAComparedtocontrols,andIL-2,IL-12,IFN-γproductioninCD4^+Tcellswasdecreased(P<0.01),IL-4andIFN-γproduc-tioninCD4^+Tcellswasnotsignificantlydifferent(P>0.05),whileIL-2andIL-12productioninCD8^+Tccellswasdecreased(P<0.01),whereasIFN-γandIL-4pro-ducinginCD4^+Tcellswereofnosignificantlydifference(P>0.05).Conclusions:TherewasanimbalanceofTlympho-cytesubsets,Th1/Th2cytokinesandTc1/Tc2intheperipheralbloodofCApatients,whichmayplayanimportantroleinthepathogenesisandprogressionofCA.

简介：obtainaninitialoverviewofgenediversityandexpressionpatterninporcinethymus,11,712ESTs(ExpressedSequenceTags)from100-day-oldporcinethymus(FTY)weresequencedand7,071cleanedESTswereusedforgeneexpressionanalysis.ClusteredbythePHRAPprogram,959contigsand3,074singletswereobtained.Blastsearchshowedthat806contigsand1,669singlets(totally5,442ESTs)hadhomologuesinGenBankand1,629ESTswerenovel.AccordingtotheGeneOntologyclassification,36.99%ESTswerecatalogedintothegeneexpressiongroup,indicatingthatalthoughthefunctionalgene(18.78%indefensegroup)ofthymusisexpressedinacertaindegree,the100-day-oldporcinethymusstillexistsinadevelopmentalstage.Comparativeanalysisshowedthatthegeneexpressionpatternofthe100-day-oldporcinethymusissimilartothatofthehumaninfantthymus.

简介：Carotenepigmentsinflowersandfruitsaredistinctfeaturesrelatedtofitnessadvantagessuchasattractinginsectsforpollinationandbirdsforseeddispersal.Inpapaya,thefleshcolorofthefruitisconsideredaqualitytraitthatcorrelateswithnutritionalvalueandislinkedtoshelf-lifeofthefruit.Toelucidatethecarotenoidbiosynthesispathwayinpapaya,wetookacandidategeneapproachtoclonethelycopeneβ-cyclasegene,LCY-B.ApapayaLCY-Bortholog,cpLCY-B,wassuccessfullyidentifiedfrombothcDNAandbacterialartificialchromosome(BAC)librariesandcompletegenomicsequencewasobtainedfromthepositiveBACincludingthepromoterregion.ThiscpLCY-Bshared80%aminoacididentitywithcitrusLCY-B.However,fullgenomicsequencesfrombothyellow-andred-fleshedpapayawereidentical.Quantitativereal-timePCR(qPCR)revealedsimilarlevelsofexpressionatsixdifferentmaturingstagesoffruitsforbothyellow-andred-fleshedgenotypes.FurtherexpressionanalysesofcpLCY-Bshowedthatitsexpressionlevelswereseven-andthree-foldhigherinleavesand,respectively,flowersthaninfruits,suggestingthatcpLCY-Bisdown-regulatedduringthefruitripeningprocess.

简介：Objective:TostudythechangesofthegeneexpressionpatternofspinalcordtissuesintheearlystageafterinjurybyDNAmicroarray(genechip).Methods:ThecontusionmodelofratspinalcordwasestablishedaccordingtoAllen'sfallingstrikemethodandthegeneexpressionpatternsofnormalandinjuredspinalcordtissueswerestudiedbygenechip.Results:Theexpressionof45geneswassignificantlychangedintheearlystageafterspinalcordinjury,inwhich22genesup-regulatedand23genesdown-regulated.Conclusions:Theexpressionofsomegeneschangessignificantlyintheearlystageafterspinalcordinjury,whichindicatesthecomplexityofsecondaryspinalcordinjury.

简介：Objective:ThepresentstudyaimedtoinvestigatecircularRNA(circRNA)expressioninuvealmelanoma(UM).Methods:First,weusedmicroarraytocomparetheexpressionprofilesofcircRNAinfiveUMsamplesandfivenormaluveatissues.Next,bioinformaticsanalyses,includinggeneontology(GO)analysisandpathwayanalysis,wereappliedtostudythesedifferentiallyexpressedcircRNAstopredictpathogenicpathwaysthatmaybeinvolved.Quantitativereal-timepolymerasechainreaction(qRT-PCR)in20UMsamplesand20normaluveasampleswasusedtoconfirmthecircRNAexpressionprofilesobtainedfromthemicroarraydata.Finally,weanalyzedtheinteractionbetweenvalidatedcircRNAsandtheirpotentialcancer-associatedmiRNAtargets.Results:Intotal,50,579circRNAs[foldchange(FC)≥2.0;P<0.05],including20,654up-regulatedand29,925down-regulatedcircRNAs,wereidentifiedasdifferentiallyexpressedbetweenUMtissuesandnormaluveatissues.WeusedqRT-PCRtoverifysevendysregulatedcircRNAsindicatedbythemicroarraydata,includinghsacirc0119873,hsacirc0128533,hsacirc0047924,hsacirc0103232,hsa-circRNA10628-6,hsacirc0032148andhsacirc0133460,whichmaybepromisingcandidatestostudyfuturemolecularmechanisms.Conclusions:Thisstudyexplored,forthefirsttime,theabnormalexpressionofcircRNAsinUManddescribedtheexpressionprofileofcircRNAs,providinganewpotentialtargetforthemechanismofUMandfuturetreatmentofUM.

简介：AbstractBackground:Recent studies have reported circular RNA (circRNA) expression profiles in various tissue types; however, circRNA expression profile in human epicardial adipose tissue (EAT) remains undefined. This work aimed to compare circRNA expression patterns in EAT between the heart failure (HF) and non-HF groups.Methods:RNA-sequencing was carried out to compare circRNA expression patterns in EAT specimens from coronary artery disease cases between the HF and non-HF groups. Quantitative real-time polymerase chain reaction was performed for validation. Comparisons of patient characteristics between the two groups were using t test, Mann-Whitney U test, and Chi-squared test.Results:A total of 141 circRNAs substantially different between the HF and non-HF groups (P < 0.05; fold change >2) were detected, including 56 up-regulated and 85 down-regulated. Among them, hsa_circ_0005565 stood out, for it had the highest fold change and was significantly increased in HF patients in quantitative real-time polymerase chain reaction validation. The top highly expressed EAT circRNAs corresponded to genes involved in cell proliferation and inflammatory response, including GSE1, RHOBTB3, HIPK3, UBXN7, PCMTD1, N4BP2L2, CFLAR, EPB41L2, FCHO2, FNDC3B, and SPECC1. The top enriched Gene Ontology term and Kyoto Encyclopedia of Genes and Genomes pathway were positive regulation of metabolic processes and insulin resistance, respectively.Conclusion:These data indicate EAT circRNAs may contribute to the pathogenesis of metabolic disorders causing HF.

简介：AbstractBackground:Hyperbaric oxygen treatment (HBOT) has been demonstrated to influence the keloid recurrence rate after surgery and to relieve keloid symptoms and other pathological processes in keloids. To explore the mechanism of the effect of HBOT on keloids, tumor immune gene expression and immune cell infiltration were studied in this work.Methods:From February 2021 to April 2021, HBOT was carried out on keloid patients four times before surgery. Keloid tissue samples were collected and divided into an HBOT group (keloid with HBOT before surgery [HK] group, n = 6) and a non-HBOT group (K group, n = 6). Tumor gene expression was analyzed with an Oncomine Immune Response Research Assay kit. Data were mined with R package. The differentially expressed genes between the groups were compared. Hub genes between the groups were determined and verified with Quantitative Real-time PCR. Immune cell infiltration was analyzed based on CIBERSORT deconvolution algorithm analysis of gene expression and verified with immunohistochemistry (IHC).Results:Inflammatory cell infiltration was reduced in the HK group. There were 178 upregulated genes and 217 downregulated genes. Ten hub genes were identified, including Integrin Subunit Alpha M (ITGAM), interleukin (IL)-4, IL-6, IL-2, Protein Tyrosine Phosphatase Receptor Type C (PTPRC), CD86, transforming growth factor (TGF), CD80, CTLA4, and IL-10. CD80, ITGAM, IL-4, and PTPRC with significantly downregulated expression were identified. IL-10 and IL-2 were upregulated in the HK group but without a significant difference. Infiltration differences of CD8 lymphocyte T cells, CD4 lymphocyte T-activated memory cells, and dendritic resting cells were identified with gene CIBERSORT deconvolution algorithm analysis. Infiltration levels of CD4 lymphocyte T cell in the HK group were significantly higher than those of the K group in IHC verification.Conclusion:HBOT affected tumor gene expression and immune cell infiltration in keloids. CD4 lymphocyte T cell, especially activated memory CD4+T, might be the key regulatory immune cell, and its related gene expression needs further study.

简介：Object:ToidentifytranscriptvariantsandexpressionpatternsofporcineMitf.Materialsandmethods:ApairwiseBLASTsearchatNCBIdatabasewasperformedtodeducethestructureofporcineMitfgene.Subsequently,50RACEandfluorescentquantitativeRT-PCRwereusedtoanalyzetheexpressionpatternofporcineMitfindifferenttissues.Results:FourtranscriptvariantsofporcineMitf,MITF-A,MITF-H,MITF-MandMITF-SUSwereidentified,allsharinghighhomologywiththoseinhumans,exceptMitf-SUS.Conclusion:ThesequenceofporcineMitfappearhighlyhomologoustohumanMITF.However,only4transcriptvariantsofporcineMitfwereidentifiedintheseminipigs,lessthanthe9transcriptvariantsinhumanMITF.

简介：WeclonedcDNAsforXenopusaldolasesA,BandC.Thesethreealdolasegenesarelocalizedondifferentchromosomesasasinglecopygene.Intheadult,thealdolaseAgeneisexpressedextensivelyinmuscletissues,whereasthealdolaseBgeneisexpressedstronglyinkidney,liver,stomachandintestine,whilethealdolaseCgeneisexpressedinbrain,heartandovary.InoocytesaldolaseAandCmRNAs,butnotaldolaseBmRNA,areextensivelytranscribed.Thus,aldolaseAandCmRNAs,butnotBmRNA,occurabundantlyineggsasmaternalmRNAs,andstrongexpressionofaldolaseBmRNAisseenonlyafterthelateneurulastage.WeconcludethataldolaseAandCmRNAsaremajoraldolasemRNAsinearlystagesofXenopusembryogenesiswhichproceedsutilizingyolkastheonlyenergysource,aldolaseBmRNA,ontheotherhand,isexpressedonlylaterindevelopmentintissueswhicharerequiredfordietaryfructosemetabolism.WealsoisolatedtheXenopusaldolaseCgenomicgene(ca.12kb)andfoundthatitspromoter(ca.2kb)containsregionsnecessaryfortissue-specificexpressionandalsoaGCrichregionwhichisessentialforbasaltranscriptionalactivity.

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简介：ToobtaintherecombinantsolubleproteinoftheextracellularfragmentofhumanTRAILgeneandtoidentifyitsfunctionpreliminarily,thisgenefragmentwasamplifiedfromperipheralbloodmononu-clearcells(PBMC)byRT-PCRandclonedintovectorpGEM-T-Easyforsequenceanalysis.Theexpres-sionvectorpET-30a/TRAILwasthenconstructedbyDNArecombinationmethodwithaHis-taggeneatthefrontoftheTRAILfragment,andtherecombinantproteinwasexpressedinE.coliBL21(DE3).Meanwhile,theexpressedtargetproteinwaspurifiedwithNi-NTAchromatographycolumnandidentifiedbySDS-PAGEandWesternblotting.TheproliferationinhibitionactivityofTRAIL-HiswasdetectedbyMTTassay.PIstainingandWright-GiemsastainingwereusedtodetectthepresenceoftheTRAIL-in-ducedcellapoptosis.ItwasdemonstratedthatthetargetproteinexpressedinE.coliBL21showedthesamerelativemolecularmassasthattheproteinexpectedandcouldberecognizedbyboththeanti-TRAILpolyclonalantibodyandanti-Hismonoclonalantibody.Inaddition,thisproteincouldalsoinhibitprolif-erationofhumanlymphomacelllineJurkatcellsorinduceapoptosisofthiscellline.ItisapparentthatarecombinantsolubleTRAILproteinwithbiologicalactivityisobtainedandthisprospectivestudycanlaysolidfoundationforfurtherresearchonthebiologicalactivityandapplicationintheanti-tumortherapy.

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简介：Sincepigisanimportantlivestockspeciesworldwide,itsgeneexpressionhasbeeninvestigatedintensively,butrarelyinbrain.Inordertostudygeneexpressionprofilesinthepigcentralnervoussystem,wesequencedandanalyzed43,122highquality5′endexpressedsequencetags(ESTs)fromporcinecerebellum,cortexcerebrum,andbrainstemcDNAlibraries,involvingseveraldifferentprenatalandpostnataldevelopmentalstages.TheinitialESTswereassembledinto16,101clustersandcomparedtoproteinandnucleicaciddatabasesinGenBank.Ofthesesequences,30.6%clustersmatchedproteindatabasesandrepresentedfunctionknownsequences;75.1%hadsignificanthitstonucleicaciddatabasesandpartialrepresentedknownfunction;73.3%matchedknownporcineESTs;and21.5%hadnomatchestoanyknownsequencesinGenBank.WeusedthecategoriesdefinedbytheGeneOntologytosurveygeneexpressionintheporcinebrain.

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简介：Inthisstudy,weexaminedtheeffectofelevatedtemperatureontheexpressionpatternsofgenes,i.e.,nacrein,irr,n16,n19,andhsp70inthepearloysterPinctadafucata.Theexperimentwascarriedoutat4temperatures,i.e.,20℃(ambient,control),24,28℃,and32℃.TheexpressionlevelsoftargetgenesinP.fucatawereassayedat0,6,24,48,and96hviareal-timepolymerasechainreaction.Resultsshowedthattheexpressionlevelsofnacreinandirrhadnosignificantvariationsamongdifferenttimepointsbelow28℃,butsignificantlyincreasedovertimeat32℃.Theexpressionlevelsofn16andn19didnotchangemarkedlyat20℃.Theformerincreasedsignificantlyat6hand24hwhilethelattersubstantiallydecreasedduring6–96hat24,28and32℃.Amongdifferenttemperatures,thelevelofn16wassignificantlylowerat20℃thanatothertemperaturesduring6–96h,andthelevelofn19significantlyvariedamongdifferenttemperaturesat48hand96h.Theexpressionlevelofhsp70wassignificantlyhigherat32℃thanat20,24and28℃at24h.TheseresultsdemonstratedthatelevatedtemperatureimpactedthephysiologicalprocessesofP.fucataandpotentiallyinfluenceditsadaptabilitytothermalstress.

简介：Objective:ToevaluatethehumoralimmuneinductioninratsofacandidateAIDSvaccineexpressingthegagp24genefromasubtypeBHIV-1isolate.Methods:Theamplifiedp24genewasinsertedintoaneukaryoticexpressionvectortoformthesupercoiledDNAvaccine.ThelinearizedexpressedDNAvaccinewaspreparedfromtheexpressionplasmidbypolymerasechainreaction(PCR).TheantigengeneexpressioninratsofthelinearizedandsupercoiledDNAvaccineswereinvitroandinvivodetected.Results:InvitrotranscriptionandNorthernhybridizationshowedthatthelinearizedDNAvaccinecouldsynthesizeamountsofp24mRNAsimilartothesupercoiledDNAvaccine.AntibodyassaysofinoculatedratsconfirmedthatthelinearizedexpressionDNAcouldinduceaslightlyhigherantibodytiterthantheexpressionplasmid,whilethehighestantibodytiterhadbeeninducedbyplasmidplusadjuvantinoculation.Conclusion:TheconstructionofacandidateAIDSvaccinebasedonthep24genecouldshedlightonapotentialHIVvaccine,meritingevaluationinarhesusmacaqueSHIV-AIDSmodel.

简介：Domesticpig(Susscrofadomestica)isoneofthemostimportantmammalstohumans.Alternativesplicingisacellularmechanismineukaryotesthatgreatlyin-creasesthediversityofgeneproducts.Expressionsequencetags(ESTs)havebeenwidelyusedforgenediscovery,expressionprofileanalysis,andalternativesplicingdetection.Inthisstudy,atotalof712,905ESTsextractedfrom101differentnon-normalizedESTlibrariesofthedomesticpigwereanalyzed.TheseESTlibrariescoverthenervoussystem,digestivesystem,immunesystem,andmeatproductionrelatedtissuesfromembryo,newborn,andadultpigs,makingcontributionstotheanalysisofalternativesplicingvariantsaswellasexpressionprofilesinvariousstagesoftissues.AmodifiedapproachwasdesignedtoclusterandassemblelargeESTdatasets,aimingtodetectalternativesplicingtogetherwithESTabundanceofeachsplicingvariant.Mucheffortsweremadetoclassifyalternativesplicingintodifferenttypesandapplydifferentfilterstoeachtypetogetmorereliableresults.Finally,atotalof1,223geneswithaverage2.8splicingvariantsweredetectedamong16,540uniquegenes.Theoverviewofexpressionprofileswouldchangewhenwetakealternativesplicingintoaccount.

简介：Objective:ToconstructtherecombinantplasmidcontainingGlycerophosphodiesterphosphodiesterase(Gpd)genefromTreponemapallidumandtransfectitintoHelacellstoexpresstheencodedoutermembraneprotein.Methods:TheGpdgenewasamplifiedfromthegenomicDNAofT.pallidumbypolymerasechainreaction(PCR)andinsertedintocloningvectorpUCm-T.TheinsertedGpdgenewassubclonedintotheappropriatesiteofpcDNA3.1(+)vector.Afteridentificationbysequencingandrestrictiveenzymesdigestion,therecombinantplasmidwastransfectedintoHelacellsusingliposomes.TheexpressedproteinwasidentifiedbyimmunocytochemistryandWesternblot.Results:ThetargetGpdgenesegmentwasapproximately1059bp.TheDNAsequenceoftheGpdgenecontainedinthepcDNA3.1(+)vectorwasconsistentwiththepublishednucleotidesequence.ThehomologyofthenucleotideandputativeaminoacidsequencesoftheGpdgenebetweenT.pallidumsubsp.pallidumNicholsandvariouspathogenictreponemalstrainsrangedfrom98%to100%.ImmunocytochemistryandWesternblotanalysisshowedthattheconstructedGpd-pcDNA3.1(+)vectorexpressedafusionproteinwithacalculatedmolecularmassof41KDainHelacellsandthattheexpressedproteinreactedwiththeserafromsyphilispatients.Conclusion:ThesuccessfulconstructionandexpressionoftheeukaryoticexpressionplasmidoftheGpdgenefromT.pallidumprovideapromisingtooltofurtherstudythebiologicalactivityofT.pallidumanddevelopaDNAvaccineforsyphilis.